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首页> 美国卫生研究院文献>Journal of Research of the National Institute of Standards and Technology >Development of Multicolor Flow Cytometry Calibration Standards: Assignment of Equivalent Reference Fluorophores (ERF) Unit
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Development of Multicolor Flow Cytometry Calibration Standards: Assignment of Equivalent Reference Fluorophores (ERF) Unit

机译：多色流式细胞仪校准标准的发展：等效参考荧光团（ERF）单元的分配

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摘要

A procedure is described for assigning the number of equivalent reference fluorophores (ERF) values to microspheres labeled with a fluorophore designed to produce a fluorescence response in a given fluorescence channel of a multicolor flow cytometer. A fluorimeter was calibrated by a series of solutions of the reference fluorophores. The fluorimeter was used to obtain the microsphere fluorescence intensity, and a multicolor flow cytometer was used to obtain the microsphere concentration. The microsphere fluorescence intensity and the concentration were used to obtain the value of ERF for each model microsphere calibration standard. The procedure is described in detail only for microspheres with allophycocyanin (APC) immobilized on the surface. ERF values were also determined for microsphere calibrators for three other fluorescence channels: fluorescein isothiocyanate (FITC), phycoerythrin (PE), and Pacific Blue(PB). The four model microsphere calibrators provide a one point calibration for the four channels of a flow cytometer. By using software controls and changing the photomultiplier voltages, it is possible to obtain a multipoint calibration for each fluorescence channel using each microsphere calibrator.
机译：描述了一种程序，该程序用于将等效参考荧光团（ERF）值的数量分配给标记有荧光团的微球，该荧光团设计为在多色流式细胞仪的给定荧光通道中产生荧光响应。通过一系列参考荧光团溶液校准荧光计。使用荧光计获得微球的荧光强度，并使用多色流式细胞仪获得微球的浓度。使用微球荧光强度和浓度获得每种模型微球校准标准品的ERF值。该过程仅针对表面固定有别藻蓝蛋白（APC）的微球进行详细描述。还为其他三个荧光通道的微球校准器确定了ERF值：异硫氰酸荧光素（FITC），藻红蛋白（PE）和太平洋蓝（PB）。四种型号的微球校准器可对流式细胞仪的四个通道进行单点校准。通过使用软件控制并更改光电倍增管电压，可以使用每个微球校准器对每个荧光通道进行多点校准。

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